Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.3.1 (alkaline phosphatase)
 47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Membrane receptors specific for IgD (IgD-R) have been identified on murine CD4+ and human CD4+ and CD8+ T lymphocytes. Up-regulation of these IgD-specific receptors can be achieved by exposure of such T cells to various stimuli, including oligomeric or Ag cross-linked IgD, IL-2, IL-4, and T cell mitogens, such as PHA. Previous studies with murine IgD-R+ splenic T cells and IgD-R+ T hybridoma cells have demonstrated the existence of soluble IgD-binding factors (IgD-BF) that are shed or released into the medium in which these cells are grown. In our study, human peripheral blood T cells and IgD-R+ T hybridoma cells were examined for their ability to produce human IgD-BF. PHA stimulation of peripheral blood T cells results in their release of an IgD-specific factor with an apparent Mr of 70 kDa. IgD- Sepharose-purified IgD-BF was able to competitively inhibit rosetting of IgD-R+ T cells with IgD-coated RBC. Immunoblot assays in which alkaline phosphatase-conjugated human IgD myeloma protein was used as a probe, confirmed the IgD specificity of IgD-BF. An IgD-BF-specific mAb (LTB9) that also reacts with membrane IgD-R was produced after immunization of BALB/c mice with this factor. LTB9 was able to detect IgD-BF in the supernatants derived from human IgD-R+, tetanus toxoid-specific T hybridoma cells, H9-CEM1, and to stain membrane IgD-R by indirect immunofluorescence. Stimulation of H9-CEM1 cells with immobilized IgD resulted in up-regulation of membrane IgD-R expression, as measured cytofluorometrically with LTB9-stained cells, and potentiated release of IgD-BF from these cells. Finally, LTB9 as well as IgD-Sepharose, immunoprecipitated a 70-kDa protein from the lysates of biosynthetically labeled H9-CEM1 cells. Similar immunoprecipitation results were obtained with H9-CEM1-derived supernatants containing IgD-BF. Taken together, these results support the hypothesis that human T cell membrane IgD-R are released as soluble IgD-BF.
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PMID:IgD-receptor-positive human T lymphocytes. II. Identification and partial characterization of human IgD-binding factor. 154 18

The study evaluates a novel version of the bromodeoxyuridine (BrdU) incorporation assay for in vitro proliferative responses, which works with finger-prick blood specimens. It was developed primarily for field-work involving children in Africa and elsewhere. Heparinized blood specimens from healthy volunteers were diluted 1:15 and cultured with purified protein derivative, tetanus toxoid, concanavalin A or medium alone for four-seven days and pulsed during the last 24 hours with bromodeoxyuridine. Thereafter, white and red cells were separated by Ficoll-Pacque centrifugation. The white cells were made to adhere to Multitest slides pre-treated with poly-L-lysin. After fixation with paraformaldehyde, the cells were incubated with mouse monoclonal antibodies to a surface marker for activated T cells (CD25, interleukin-2 receptor) and the reaction visualized with anti-mouse alkaline phosphatase conjugated antibodies and a Fast Red substrate. After treatment with cold acetone, the cells were covered with formamide and heated to 70 degrees C, without loss of surface staining. The incorporation of bromodeoxyuridine was visualized in the UV-microscope with mouse anti-bromodeoxyuridine monoclonal antibodies and a rabbit anti-mouse fluorescein conjugate. Both the surface staining and the nuclear fluorescence can be seen in UV-light and the cells be classified as single- or double-positive or negative. Parallel experiments on the same blood-sample from seven donors allowed for at least 42 paired comparisons between the proportion of BrdU positive cells and thymidine incorporation stimulation indices. The latter assay was carried out both in the conventional version and in a whole-blood micro-version. The rank order correlations were 0.84 and 0.91 respectively.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Whole-blood microassay for immunodetection of antigen specific cell mediated immunity using bromodeoxyuridine incorporation. 166 23

Heterotopic ossification is a very rare complication of tetanus. In one case heterotopic ossification occurred around both hip joints. The values of serum creatine phosphokinase had been elevated significantly when the patient had major muscle spasms. The elevation in serum alkaline phosphatase values following that in creatine phosphokinase values persisted for about four weeks. Partial resection of the bone mass about the right hip joint resulted in a satisfactory improvement in performance of daily activities. The specimens incised at operation revealed both lamellar and woven bone surrounded by fibrous connective tissue. Very near the bone mass were severely degenerated muscle fibers. In addition, evidence suggesting metaplasia of fibroblasts to osteoblasts was seen in some areas. Clinical and laboratory data indicate trauma as a main etiologic factor of heterotopic ossification following tetanus. Heterotopic bone formation should be considered if elevation of the serum alkaline phosphatase values persists beyond the period of elevation of serum creatine phosphokinase values. Artificial ventilation may be beneficial for preventing heterotopic ossification following tetanus if it is administered before there is significant elevation of the serum creatine phosphokinase values.
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PMID:Heterotopic ossification of the hip as a complication of tetanus. 680 96

The activities of leukocyte alkaline phosphatase (l-AP) and serum gamma-glutamil transferase (gamma-GT), and total leukocyte counts were determined in horses submitted to the production of hyper immune sera against tetanus (Clostridium tetani). The purpose of this work was to investigate the prospective changes of mentioned parameters in horses under the described circumstances. In addition, the suitability of these parameters in assessing the health condition of the same horses had to be evaluated. The average total leukocyte count increased in one month from the values considered as physiological (8.8 x 10(9)/L, Schalm et al. 1975) to the value of 12 x 10(9)/L and remained at this level up to the end of the trial. The I-AP activities fell after 30 days of the trial for 100 units, and showed permanent slight decreasing tendency thereafter. On the contrary, the serum gamma-GT activity was increasing gradually throughout the whole trial. The results indicate the possibility of reflecting the dynamics of intensified leukocyte metabolism in the course of its function within the chronic inflammation, and development of accompanying pathological changes in the liver. In addition, the initialisation period of pathological changes in the organism of horses in experiment could be between the fourth and sixth month following exposure to antigen.
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PMID:Leukocyte alkaline phosphatase and serum gamma-glutamil transferase activities in horses used for production of hyper immune sera. 750 8

1. Endosulfan insecticide is a polychlorinated compound used for controlling a variety of insects; it is practically water-insoluble, but readily adheres to clay particles and persists in soil and water for several years. Its mode of action involves repetitive nerve-discharges positively correlated to increase in temperature. This compound is extremely toxic to most fish and can cause massive mortalities. In fish, it causes marked changes in Na and K concentrations, decrease in blood Ca(2+) and Mg levels and inhibits Na, K and Mg-dependent ATPase (in brain). 2. Bioaccumulation of endosulfan is reported for marine animals; however, freshwater animals (e.g., crayfish) accumulate it to some extent, but they lose the compound rapidly during depuration. Endosulfan is generally less toxic to aquatic invertebrates than fish. However, it causes decreases in adenylate energy charge, oxygen consumption, hemolymph amino acids, succinate dehydrogenase, heart-beat (mussel) and altered osmoregulation. 3. Generally, mammals are less susceptible to endosulfan's toxicity than aquatic animals. The majority of studies conducted on laboratory mammals can be summarized. (a) Neurotoxicity: male rats are more sensitive than females to endosulfan, which decreases brain and plasma acetylcholinesterase activity. Endosulfan I (a metabolite) causes a significant change in norepinephrine, 5-HT and GABA. (b) Renal toxicity: inhibition of MFOs activity was noticed in rats; other effects included changes in proximal convoluted tubules and necrosis of the tubular epithelium. (c) Hepatotoxicity: chemically-induced aminopyrine N-demethylase and aniline hydrolase were found in rat liver, and reduction in the glycogen level occurred. (d) Hematologic toxicity: endosulfan exposure resulted in a significant decrease in the level occurred. (d) Hematologic toxicity: endosulfan exposure resulted in a significant decrease in the erythrocyte glutathione reductase, hemoglobin amount, RBC number and mean corpuscular volume. 4. Respiratory toxicity: involved dyspnea, acute emphysema, cyanosis and hemorrhages in teh interalveolar portions of rat's lungs. 5. Biochemical: in rats, endosulfan caused increased glucose-6-phosphate dehydrogenase activity, blood glucose level, phospholipid contents of the microsomal and surfactant system, and profoundly induced the activity of alcohol dehydrogenase and cytosolic glutathione S-transferases. It also decreased significantly Na+, K+ and Mg(2+) ATPases, plasma calcium level and alkaline phosphatase in the intestinal epithelium. 6. Immunologic toxicity: rat serum antibody titer to tetanus toxin, IgG, IgM and gammaglobulins were significantly reduced. 7. Reproductive toxicity: degenerative changes in the seminiferous epithelium, induction of the rate-limiting enzyme in testosterone production (3beta-hydroxysteroid transferase and 17 beta-hydroxysteroid transferase), histological changes in reproductive organs, testicular atrophy and the occurrence of ovarian cysts were noticed in rat. Reduction in the weight of secondary sex organ was also observed.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Bioaccumulative potential and toxicity of endosulfan insecticide to non-target animals. 790 Sep 59

The purpose of this study was to compare, using cell blot analysis, the association of gingival tissue mononuclear cells (GTMC) isolated from lesions displaying histories of early-onset periodontitis (EOP; typically B-lymphocyte dominated) and gingivitis (typically T-lymphocyte dominated) with the B-cell stimulating cytokine, interleukin (IL)-4, and the T-cell stimulating cytokine, IL-2. Eleven EOP patients and 11 age- and gender-similar gingivitis control (GC) subjects participated. Gingival tissue adjacent to the alveolar crest normally removed during surgery was digested in collagenase-containing media and GTMC were isolated by density gradient centrifugation. Cells were separated into four aliquots. One was left unstimulated; the remainder were stimulated for 2 hours with Porphyromonas gingivalis outer membrane protein, mitogen Concanavalin A, or common antigen tetanus toxoid. Cells then were centrifuged onto transfer membranes and incubated in RPMI 1640 media for 6 hours to allow absorption of secreted cytokine. Membranes were treated with monoclonal anti-IL-2 or anti-IL-4, followed by a biotin-conjugated second layer, streptavidin-alkaline phosphatase and nitro blue tetrazolium/5-bromo-4-chloro-indolyl-phosphate (NBT/BCIP) color development. A higher percentage of GTMC from EOP patients were IL-2+ when stimulated with P. gingivalis compared with GTMC from GC patients (20 +/- 2% vs. 12 +/- 2%, P < 0.003). A higher percentage of non-stimulated GTMC from EOP patients produced IL-4 than from GC (22 +/- 4% vs. 6 +/- 3%, P < 0.00007), as well as when stimulated with P. gingivalis (22 +/- 3% vs. 13 +/- 2%, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Gingival cell IL-2 and IL-4 in early-onset periodontitis. 799 15

A modified fast micro method for spectrotypic/clonotypic analysis of human IgG1-4 antibodies against bacterial virulence antigens of polysaccharide or protein nature is described. Serum samples of as small volumes as 0.5 microliter were isoelectrically focused in micro agarose gels made in plexiglass matrices and blotted using immunoaffinity-mediated capillary blotting onto nitrocellulose membranes previously coated with antigen. The bands of the antigen-specific antibodies were identified with respect to isotypes, light chain types, allotypes or idiotypes by incubating the nitrocellulose membranes with mouse monoclonal anti-human IgG subclass antisera and then with alkaline phosphatase-conjugated rabbit anti-mouse immunoglobulins. The method was applied for characterization of human monoclonals against tetanus toxoid (TT) and for the analysis of variable patterns of clonotypes in IgG subclass-deficient patients. The usefulness of the technique was also demonstrated by comparing the variable specificity and reactivity of different commercial monoclonals against human IgG subclasses. This method is fast, specific, sensitive, uses little material, is simple and reproducible.
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PMID:A modified fast micro method in agarose for isotype, allotype, light chain and idiotype-specific analysis of antibody clonotypes to bacterial virulence antigens. 1008 3

The importance of conserved inner saccharide epitopes to the immune performance of meningococcal lipooligosaccharide-protein conjugate vaccines was demonstrated in the following experiments. Two different oligosaccharides were obtained by chemical degradations of the same L7 lipooligosaccharide, and both were linked terminally to tetanus toxoid. One was a truncated oligosaccharide in which the inner epitopes were incomplete and was obtained by mild acid hydrolysis of the L7 lipooligosaccharide. This oligosaccharide was conjugated by direct reductive amination through its newly exposed terminal Kdo residue. The second, a full-length oligosaccharide, was obtained by O-deacylation of the L7 lipooligosaccharide, with subsequent removal of phosphate substituents from its lipid A moiety using alkaline phosphatase. This permitted the full-length oligosaccharide to be conjugated directly to tetanus toxoid by reductive amination through its newly exposed terminal 2-N-acyl-2-deoxy-D-glucopyranose residue. Comparison of the immune performance of the two conjugates in mice revealed, that while both were able to induce significant levels of L7-lipooligosaccharide-specific IgG antibody, the conjugate made with the full-length saccharide was able to induce antibodies with increased bactericidal activity against homologous meningococci.
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PMID:Conjugation of meningococcal lipooligosaccharides through their lipid A terminus conserves their inner epitopes and results in conjugate vaccines having improved immunological properties. 1252 40

Cationization is a strategy to enhance the permeability of antibodies to physiological membranes for potential therapeutic and diagnostic applications of these proteins, with one of its crucial points being the retention of antigen binding activity. Here, we describe the cationization of horse polyclonal anti-tetanus F(ab')(2) fragments and the development and validation of an ELISA for quantitative measurements of the binding activity of the native and cationized F(ab')(2) in cell lysates and rat plasma samples, assessing the cellular uptake and plasma kinetics of these antibodies, respectively. The method used tetanus anatoxin coated on microtitre plates as capture antigen to bind sample or standard F(ab')(2), the amount of antibody binding being quantified using, first, a secondary biotinylated anti-horse antibody/streptavidin-alkaline phosphatase complex in situ and then a measurement of the substrate product. Cationization of the F(ab')(2) was performed with putrescine at pH 4.5 using soluble carbodiimide as carboxyl activator. The average substitution ratio was determined at 3 putrescine molecules per F(ab')(2) molecule. The cationized F(ab')(2) retained roughly 80% of the initial antigen binding activity and was stable over a 1 year period of storage at -20 degrees C. The ELISA validation data showed that the method was linear for both the native and cationized F(ab')(2) using Hanks' balanced saline solution with 0.2% bovine serum albumin as assay diluent for the cell lysate samples. The useful F(ab')(2) concentration range was 2.5-25 ng/ml and the limit of quantification was 2.5 ng/ml. With rat blank plasma used as assay diluent for the rat plasma samples the useful F(ab')(2) concentration range was 3.5-25 ng/ml and the limit of quantification was 3.5 ng/ml. Specific requirements for the limits of quantification were fulfilled: precision < or =20% CV and accuracy within +/-20% of the nominal values. Intra- and inter-assay coefficients of variation were within 9% and accuracies within +/-10% of the nominal values. The validated method was applied to the study of the cellular uptake of native and cationized anti-tetanus F(ab')(2) in an HL 60 cell model, and of plasma kinetics after i.v. administration to rats.
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PMID:Development and validation of an antigen-binding capture ELISA for native and putrescine-modified anti-tetanus F(ab')2 fragments for the assessment of the cellular uptake and plasma kinetics of the antibodies. 1630 97